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PubMed1979

Ultrastructural changes in cells associated with interkinetic nuclear migration in the developing chick neuroepithelium.

Changes in fine structure of cells associated with interkinetic nuclear migration in the developing chick neuroepithelium were investigated. Interphase cells are elongated and span the entire thickness of the neuroepithelium. As cells round up in preparation for mitosis, they sever their contacts with the basement membrane, but retain their apical junctions. Meanwhile, microtubules lose their apico-basal orientation and the apical microfilament bundle relaxes to allow broadening of the luminal surface. These changes in the cytoarchitecture together with an increased cytoplasmic viscosity may cause rounding of mitotic cells and their juxtaluminal position. Mitotic cells remain at the lumen from late prophase through early telophase. By late telophase, daughter cells start to elongate toward the base of the neuroepithelium. The ultrastructural changes during elongation recapitulate, in a reverse order, the events of rounding up in preparation for mitosis. Daughter cells are connected for some time after mitosis by a thread of cytoplasm. The thread is filled with microtubules representing a remnant of the spindle complex and has an electron-dense midbody at about the middle of its length. During the final stage of separation of daughter cells, the thread is split at the level of the midbody.

Animals↗
PubMed1982

Studies of mitosis in excised limb regenerates of the newt, Notophthalmus viridescens.

Subsequent to excision and explantation of the limb blastema into culture medium, there is an abrupt reduction in mitotic index lasting several hours. Coincident with the disappearance of mitosis, abnormal mitotic figures (AMFs), lacking the condensed chromosomal nature of normal figures, are seen. These persist until normal levels of mitotic activity are restored approximately 6 hr later. The transient loss of mitotic activity is observed in both ganglionated and nonganglionated explants. The large number of prophase figures observed at time zero were sharply reduced within minutes, without concomitant increases in the later phases. The possibility that some cells, without completing mitosis, become temporarily indistinguishable as mitotic figures, is discussed in terms of chromosomal decondensation and recondensation.

Animals↗
PubMed1983

Syntheses of nucleic acids during spermatogenesis in Nereis diversicolor (annelida polychaeta): a quantitative autoradiographic study.

The development of DNA and RNA synthesis in the germ cell population was studied after a 3H-thymidine or 3H-uridine pulse at each stage of spermatogenesis. The autoradiographic results show that the first sign (after 3 days in vitro) of cellular changes is an increase in RNA synthesis which reaches a maximum at day 5. DNA replication (premeiotic S phase) occurred at day 7, then cells entered meiotic prophase (day 9). Meiotic divisions and spermiogenesis occurred after 11 days. Silver grain counts permit the conclusion that RNA synthesis is clearly higher during premeiotic interphase (days 3-7) than during spermatogonial proliferation (day 0). It appears therefore that male meiotic differentiation in Nereidae is accompanied by increased RNA synthesis.

Animals↗
PubMed1983

Hydrostatic pressure inhibition of hormone-induced resumption of meiotic maturation in isolated oocytes.

Prior to ovulation, fully grown oocytes of both the amphibian (Rana pipiens) and the starfish (Piaster ochraceus), like those of many other organisms, are arrested in late prophase I of meiosis. Reinitiation of meiotic maturation in oocytes from either of these organisms is hormonally induced. Although the meiosis-inducing substance (MIS) for each organism is chemically dissimilar (a steroid in the frog and a purine in the starfish) induction of oocyte maturation in both biological systems appears to be initiated by the interaction of the MIS with the plasma membrane of the oocyte. The objective of this investigation was to determine if elevated hydrostatic pressure affected hormonal induction of oocyte maturation and to compare the effect of pressure on oocytes stimulated with different meiosis-inducing substances. In isolated oocytes from either the frog or the starfish, increasing ambient pressure reduced the percentage of oocytes which matured, and a stepwise increase in pressure resulted in a corresponding shift in the dose response curves of hormone-induced, oocyte maturation to the right. In experiments using only starfish oocytes, this inhibitory effect of pressure was found to be reversible and limited to the initial period of the maturation event. Taken together, these data suggest that elevated ambient pressure inhibits an early cellular event in the hormonal induction of meiotic maturation which is common to both amphibian and starfish oocytes.

Adenine↗
PubMed1983

Germ cell differentiation in mouse adrenal glands.

The differentiation of germ cells in the adrenal glands of 26 male and female Swiss albino mice was studied in sequential stages of development, from day 12 1/2 of intrauterine life to postnatal day 21; the study was performed by means of high-resolution light microscopy and electron microscopy. In 12 1/2- and 13-day-old embryos, the ectopic cells had morphologic characteristics typical of primordial germ cells, whereas in 14- and 15-day-old fetuses they were identifiable as oogonia. In male and female fetuses from day 17 to term, all ectopic germinal elements entered meiotic prophase, reached diplotene, and differentiated into oocytes in perfect adherence to mouse ovarian timetables. In the postnatal animals, females as well as males, all oocytes progressed through the postmeiotic phase of growth just as they normally do in ovarian follicles, and, in the 2- and 3-week-old animals, they displayed features identical to those exhibited by oocytes in large antral follicles, including a zona pellucida. Germinal elements were no longer seen in the adrenals of animals older than 3 weeks. Our study shows that mammalian germ cells are capable of developing even outside the gonads, and that in ectopic sites they all differentiate as oocytes irrespective of their genetic sex.

Adrenal Cortex↗
PubMed1983

Developmental aspects of X chromosome inactivation in eutherian and metatherian mammals.

The single active X principle has served for two decades as a focal point for research on the cyclic activation and inactivation of gene loci. Differences in X chromosome inactivation patterns of eutherian and marsupial mammals provide probes for investigating the mechanisms of the X inactivation process. In eutherian mammals, the X chromosome is inactivated early in meiotic prophase in males and remains inactive throughout the rest of spermatogenesis. During meiosis in females, the inactive X chromosome is activated so that both X chromosomes are active in oocytes. During the early cleavage divisions of female embryos, the paternally derived X is activated. It and the maternally derived X remain active until differentiation begins in early embryogenesis. At that time, the paternally derived X is inactivated in cells that give rise to extraembryonic membranes, whereas a random process determines which X chromosome is inactivated in cells that give rise to the embryo itself. Although less is known about developmental aspects of X inactivation in female marsupials, it is clear that the paternal X is preferentially inactive in postembryonic somatic cells. Furthermore, the paternal X is partially active at some loci in some cell types, indicating that it is not regulated as a single unit. The successful adaptation of a small (80-150 g), fecund marsupial to simple laboratory conditions now enables extensive experimentation on the large number of marsupials at various developmental stages. This capability, coupled with the application of newly developed cellular and molecular techniques to questions about X chromosome inactivation, shows great promise for advancing our understanding of the mechanisms that control the cyclic behavior of X chromosome activity.

Animals↗
PubMed1985

Differentiation of mouse ectopic germinal cells in intra- and perigonadal locations.

Four hundred and thirteen ectopic germinal cells in the testicular and extratesticular stroma and in the rete testis of mouse fetuses from day 13 of uterine development to term were studied together with 161 ectopic germinal cells in the rete ovarii and periovarian stroma of female fetuses at days 17 and 18 of intrauterine life. The morphology and the differentiation of these ectopic germinal cells were compared to those of germinal cells within seminiferous and ovigerous cords. While the ectopic germinal cells in the testis and in the rete testis followed patterns of differentiation identical with those in the seminiferous cords throughout the period included in the study, those in the extratesticular stroma behaved like entopic germinal cells only through day 17, since at days 18 and 19 many of them entered meiotic prophase just like XX germinal cells in the ovigerous cords. No differences were noted between ectopic and entopic ovarian germinal cells. The results of this study show that the factors responsible for the male differentiation of XY germinal cells are not limited to the seminiferous cords but operate throughout the testicular territory, and confirm that outside the testis, XY germinal cells differentiate as female; our study also corroborates the thesis that the differentiation of XX germinal cells is an autonomous and ubiquitous process.

Animals↗
PubMed1987

Actin-plasma membrane associations in mouse eggs and oocytes.

Using rhodamine-phalloidin stained preparations and extracted specimens labeled with heavy meromyosin or run on polyacrylamide gels, actin-plasma membrane associations in mouse mature eggs at the second metaphase of meiosis and oocytes at meiotic prophase have been examined. Cortices of extracted oocytes possessed numerous actin filaments that emanated from the plasma membrane delimiting regions between microvilli and from microvillar apices. The membrane anchorage sites of actin filaments were marked by an electron dense material on the inner leaflet of the plasma membrane. The free ends of filaments emanating from the plasma membrane of oocytes intermeshed to form a dense, cortical layer. With meiotic maturation, changes in the organization of cortical actin were first noted approximately 3 hr after the chromosomes had become localized at the oocyte's periphery. Fewer and shorter actin filaments, which did not form a well-defined layer as in oocytes, were connected with electron-dense material to the inner leaflet of the plasma membrane of extracted egg cortices in regions other than that associated with the meiotic spindle. Cortical actin adjacent to the meiotic spindle, however, was organized into a dense, cresentic aggregation in which clusters of filaments emanated from electron-dense regions associated with both the inner and outer leaflets of the plasma membrane. These observations indicate that mouse oocyte maturation not only involves changes in the distribution of cortical actin but also local alterations in the association of actin with the plasma membrane.

Actins↗
PubMed1988

Formation of flagella during interphase in secondary spermatocytes from Xenopus laevis in vitro.

In cell culture, single motile flagella, 1 micron in length, were observed to grow from secondary spermatocytes of Xenopus laevis within 2-3 hours after telophase I, at 22 degrees C. About 90% of the secondary spermatocytes formed flagella as observed by phase-contrast microscopy. The flagella grew up to 2-6 microns in length during interphase II, which lasted about 18 hours. The presence of the "9 + 2" microtubular structure of the flagellar axonemes of secondary spermatocytes was confirmed by electron microscopy. When chromosomal condensation began (prophase II), the flagella were resorbed into the cells and, after the second meiotic division, a flagellum was formed again by each of the round spermatids. Thus, there appears to be a close relationship between the meiotic division cycle and the formation of flagella. The possible contribution of Sertoli cells to the formation of flagella in secondary spermatocytes was examined by reducing the number of Sertoli cells to less than ten per culture. Under these conditions, flagella formed in secondary spermatocytes with very high efficiency. It is very likely that secondary spermatocytes form flagella in vivo, since the secondary spermatocytes were observed to have flagella immediately after dissociation of the testes.

Animals↗
PubMed1992

Sexually differentiated mechanisms of sterility in interspecific hybrids between Oryzias latipes and O. curvinotus.

Fertility of interspecific hybrids between Oryzias latipes and O. curvinotus was examined. F1 females were able to lay eggs but males were sterile. Histological examination of the ovaries of hybrids revealed that oogenesis does not proceed normally in spite of the apparent fertility. Most oocytes degenerated at the pachytene stage of the meiotic prophase, and only a few entered the diplotene stage to develop into ova. Hybrid males could induce females to spawn eggs, an indication that they had differentiated completely into true males. However, they did not produce fertile sperm. Most germ cells in testes of hybrids passes through almost the entire process of spermatogenesis, but deviations from the normal course of events were observed during spermiogenesis. The condensation of chromatin in spermatids occurred, but the diameters of sperm heads were about 1.5-fold larger than those of normal ones. Prominent abnormalities were apparent in the quantity and arrangement of microtubules in the cytoplasm. Abnormal spermatozoa were phagocytized by Sertoli cells. These observations indicate that the mechanisms of impaired gametogenesis in these interspecific hybrids are sexually differentiated.

Animals↗
PubMed1993

Progestins inhibit murine oocyte meiotic maturation in vitro.

The in vitro culture of fully grown mammalian oocytes results in spontaneous meiotic maturation from prophase arrest to metaphase II. This maturation can be inhibited by steroid hormones in both murine and porcine oocytes. Using selected steroids, we have examined the structure-activity relationships of steroids and oocyte inhibition. Experiments with androgens, estrogens, glucocorticoids, and progesterone revealed that at least one steroid from each class was inhibitory. Progesterone, however, was two to three times more effective than steroids from other classes. Examination of a variety of progestins showed that most substitutions decreased or abolished the inhibitory activity. Hydroxy group substitutions at different carbon atoms and substitutions at the 4-ene group lessened the inhibitory effectiveness, with the exception of 5 beta-dihydroprogesterone, which was as effective as progesterone. However, several steroids with substitutions at the C17 acetyl group were more active than progesterone, including 20 beta-dihydroprogesterone which was the most inhibitory steroid tested (ID50 = 5 microM). The progesterone agonist R5020 was also very active (ID50 = 8 microM). This is the first report of a detailed examination of the steroid-induced inhibition of murine oocytes. A comparison between the results reported here and previous reports of steroid-induced inhibition in porcine oocytes reveals differences in the response of oocytes from the two families. The structure-activity relationships of the inhibitory steroids examined here suggest that the steroids are acting via a receptor-mediated system.

Animals↗
PubMed1994

In vivo and in vitro induction of germinal vesicle breakdown in a freshwater bivalve, the zebra mussel Dreissena polymorpha (Pallas).

Oocyte maturation and germinal vesicle breakdown (GVBD) was induced in zebra mussel (Dreissena polymorpha) oocytes by in vivo and in vitro application of serotonin (5-hydroxytryptamine, 5-HT), and in vitro application of 8-hydroxydipropylaminotetralin hydrobromide (8-OH-DPAT, a 5-HT1A receptor agonist). Oocytes initiated GVBD approximately 30 minutes after exposure to 5-HT (10(-3) M) at 23 degrees C, and by 40-50 minutes after exposure, most oocytes lacked a germinal vesicle. An exposure time to 5-HT as brief as five to ten minutes was required to trigger the maturation process, which terminates in spawning of fertilizable oocytes in nearly all mussels. But, with an exposure time of less than five minutes, spawning was reduced by application of 10(-4) M methiothepin (a potent blocker of 5-HT-induced spawning in zebra mussels). Thus, the sequence of oocyte maturation events in zebra mussels was determined. Oocytes are arrested at the germinal vesicle stage (prophase I) within the ovary. 5-HT reinitiates the maturation process, including GVBD and spawning of metaphase I oocytes, which are further arrested until fertilization. To our knowledge this is the first demonstration of oocyte maturation induction by serotonergic ligands in a freshwater bivalve.

8-Hydroxy-2-(di-n-propylamino)tetralin↗
PubMed1977

Spermatogenesis and spermiogenesis in Ascaris lumbricoides Var. suum.

Reorganization of the prophase I nucleus marks the beginning of the first meiotic division. A pair of centrioles is present at each pole at metaphase I and mitochondria are not observed in the spindle area. A chromosomal pellicle, which resembles a kinetochore plate but has no apparent association with microtubules, surrounds each autosome at metaphase I and II. The sex body lags behind the autosomes at anaphase I and segregates differentially to one daughter cell. Mitochondria and a pair of centrioles are present in the spindle during the second meiotic division. Localized condensation of chromatin and fusion of the condensed chromatin of the secondary spermatocyte telophase nucleus results in a compact spermatid nucleus. Loss of spermatid cytoplasm is effected by the ejection of a cytophore vesicle.

Animals↗
PubMed1983

The ultrastructure of germinal beds in the ovary of Gerrhonotus coeruleus (Reptilia: Anguidae).

A study of ovarian structure in adult Alligator Lizards (Gerrhonotus coeruleus) was conducted by light microscopy and transmission electron microscopy. Particular attention was directed to characterizing the ultrastructure of germ-line cells, prior to follicle formation. General ovarian structure in this lizard is similar to that of other lizards. The paired organs are hollow, thin-walled sacs containing follicles in roughly 3 to 4 size classes. Ovarian germinal tissue consists of oogonia (diploid cells which divide mitotically) and oocytes (meiotic cells), intermixed with ovarian surface epithelial cells. Germ cells reside in two dorsal patches of epithelium per ovary (germinal beds), as is common in lizards. Oogonia in interphase show a highly dispersed chromatin pattern. Within oogonia cytoplasm, Golgi complexes are scarce, rough endoplasmic reticulum is absent, and lipid droplets are rare. Ribosomes are scattered in small clusters. Small, round vesicles are common in all oogonia; glycogen-like granules are present in some. Mitochondria form a juxta-nuclear mass within which groups of several mitochondria surround a dense granule. 'Nuage' granules also are found unassociated with mitochondria. Oocytes are present in stages of meiotic prophase up to diplotene. Synaptinemal complexes are seen in several (pachytene) cells. The cytoplasm of oocytes differs from that of oogonia in that mitochondria do not form groups, and nuage and glycogen are absent, whereas small round vesicles and large irregular vesicles are common. The ultrastructural similarities in germ cells of a reptile as compared to those of other vertebrates strengthens the notion that germ-line cells possess (or lack) qualities related to the undifferentiated state of these cells.

Animals↗
PubMed1990

Maturation-associated changes in the rat zona pellucida.

Rat follicular oocytes, arrested at prophase I, cannot be fertilized in vitro. This capacity is acquired following resumption of meiosis and a series of changes involving both the oocyte and the cumulus cells surrounding it. Oocytes exposed to sperm at different hours before ovulation show a gradual increase in the permeability of their zona pellucida (ZP). Our study examined whether the ZP, in response to the physiological stimulus for maturation and concomitant with the other oocyte--cumulus components, undergoes maturational changes. Two ZP characteristics were assessed, sensitivity to proteolysis and sperm binding. ZP surrounding oocytes and eggs were collected from five sources: 1) germinal vesicle (GV)-intact oocytes, 2) preovulatory eggs, 3) ovulated eggs isolated from oviducts of immature females, 4) fertilized eggs, 5) ovulated eggs isolated from oviducts of mature females. All ZP surrounding oocytes/eggs from groups 1-5 were dissolved by trypsin. When solubility by pronase and alpha-chymotrypsin was examined, a large variation between groups was found. All ZP from group 2 were dissolved by 0.001% pronase, compared to 0% solubility in group 4. Only 10% of the ZP surrounding GV-intact oocytes (group 1) were dissolved by this enzyme, compared to 82% in group 3. Solubility in 0.01% alpha-chymotrypsin showed a similar pattern. Capacitated sperm were incubated with eggs from groups 1 and 3. The number of sperm binding to ZP in group 3 was repeatedly higher than that in group 1. In both tests it was found that the ZP surrounding the mature eggs differ in their characteristics from ZP of GV-intact oocytes.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1990

Stage-specific expression of phosphoprotein p19 during spermatogenesis in the rat.

The expression of phosphoprotein p19, a 19-kDa cytosolic substrate for cyclic adenosine monophosphate (cAMP)-dependent protein kinase, occurs abundantly in brain and testis and is developmentally regulated. In the present study we have identified the cell types of adult rat testis that contain p19. Using cryostat sections, which were first incubated with rabbit anti-p19 for immunohistochemistry followed by counterstaining with periodic acid-Schiff (PAS)-hematoxylin to reveal nuclear morphology, we demonstrate that immunoreactive p19 is detectable only in germ cells and is restricted to a limited stage of spermatogenesis. Expression first appears after the differentiating gametes have entered the prophase of meiosis, is abundant in spermatocytes until meiosis is completed, and declines to undetectable levels in maturing spermatids. We have ruled out immunocross-reactivity with SCG10, a 22-kDa protein that is closely related in structure to p19, by demonstrating, using Northern blot analysis, that RNA transcripts encoding SCG10 are not detectable in adult rat testis, whereas p19 is abundantly expressed. The transient expression of p19 during spermatogenesis suggests that the protein plays a role during male gamete differentiation.

Animals↗
PubMed1991

Stage and lineage-regulated expression of two hsp90 transcripts during mouse germ cell differentiation and embryogenesis.

The expression of members of the heat shock protein 90 (hsp90) gene family during testicular and embryonic development was investigated. Two different hsp90 transcripts were detected in RNA from mouse testis, approximately 3.2 kb and 2.9 kb in size, and were shown to exhibit cellular and developmental stage specificity of expression. The larger, more abundant transcript was expressed at high levels in the germinal compartment of the testis, particularly in germ cells in meiotic prophase. The smaller hsp90 transcript was expressed predominantly in the somatic compartment of the testis. Expression of the two hsp90 transcripts was observed in testes of other species, suggesting an important role for hsp90 in mammalian testicular function. In addition, expression of both hsp90 transcripts was detected in the embryonic and extra-embryonic compartments of mid-gestation embryos.

Animals↗
PubMed1991

Spermatogenesis in XY, XYSxra and XOSxra mice: a quantitative analysis of spermatogenesis throughout puberty.

Adult XYSxra mice exhibit varying degrees of spermatogenic deficiency but are usually fertile, while XOSxra mice have severe spermatogenic failure and are always sterile. The present quantitative spermatogenic analysis documents when these anomalies first appear during puberty. The results demonstrate that in XYSxra mice there was increased degeneration of pachytene spermatocytes and, to a lesser extent, meiotic metaphase stages. On average, there were only one-half the number of spermatids compared with the XY controls. The defect in XOSxra mice appeared a little later, with an almost complete arrest and degeneration during the meiotic metaphases, so that the number of spermatids produced was only 3% of the control value. These results are discussed in relation to an hypothesis that links sex chromosome univalence during meiotic prophase with spermatogenic failure.

Animals↗